2 replies to this topic

[metta]

Hi,
i'm newbie for cell culture. today when i was preparing mouse embryonic fibroblasts, after mincing the embryo, by mistake instead of trypsin i let the tissue in pensillin+strepto+glutamin for 10 min. once i realized my mistake, i centrifuged it briefly and then washed in PBS and then suspended it  in trypsin. My question is, will this mistake be a problem or its ok.
another thing is after how much time i can see fibroblasts adhering to the surface?

thanks in advance
m

[cellcounter]

metta, on Feb 24 2009, 07:33 AM, said:

Hi,
i'm newbie for cell culture. today when i was preparing mouse embryonic fibroblasts, after mincing the embryo, by mistake instead of trypsin i let the tissue in pensillin+strepto+glutamin for 10 min. once i realized my mistake, i centrifuged it briefly and then washed in PBS and then suspended it  in trypsin. My question is, will this mistake be a problem or its ok.
another thing is after how much time i can see fibroblasts adhering to the surface?

thanks in advance
m

MEFs are robust cells, so I don't see any problem with the time lapse. I would however worry if your antibiotics were 100X conc.

You will anyway see if MEFs are growing the next day. So, by now you should know the answer.

[sarigsk]

fibroblasts adhere within 30-35 min on gelatinized plates and this method is usually used to remove MEF feeder cells from co-culture with stem cells